It is known that more than 200 animal fungal pathogens and more than 30 common plant fungal pathogens can make great impact on human health and economics. At present, the major drugs for controlling human fungal pathogens are the small molecules such as polyenes, azoles, fluconazole, amphotericin B, and the like. With the increasing number of drug abuse, the situation of drug resistance of the fungal strains becomes more and more severe. There is an urgent need of developing new antifungal drugs (see Selitrennikoff, C. P., 2001, Antigungal proteins, Appl. Environ. Microbiol. 67, 2883-2894, and Liu Y., Ryan, M. E., Lee, H. M., Simon, S., Tortora, G., Lauzon, C., Leung, M. K. and Golub, L. M., 2002, A chemically modified tetracycline (CMT-3) is a new antifungal agent. Antimicrob. Agents Chemother. 46, 1447-1454).
Plants, bacteria, fungi, insects, birds and mammals are all known to be able to produce antifungal proteins (see Kaiserer, L., Oberparieiter, C., Weiler-Görz, R. Burgstaller, W., Leiter, E. and Marx, F., 2003, Characterization of the Penicillium chrysogenum antifungal protein PAF. Arch Microbiol. 180, 204-210). Although these proteins have different amino acid sequence and quaternary structure, their properties of low molecular weight, highly basic and high cysteine content are the major molecular characteristics for most antifungal proteins (see Selitrennikoff et al., 2001).
There are a few antifungal proteins from filamentous fungi been studied, for example AFP protein from Aspergillus giganteus (see Wnendt, S., Ulbrich, N. and Stahl, U., 1990, Cloning and nucleotide sequence of a cDNA encoding the antifungal-protein of Aspergillus giganteus and preliminary characterization of the native gene. Nucleic Acids Res. 18, 3987, Wnendt, S., Ulbrich, N. and Stahl, U., 1994, Molecular cloning, sequence analysis and expression of the gene encoding an antifungal protein from Aspergillus giganteus. Curr. Genet. 25, 519-523, Theis, t., Marx, F., Salvenmoser, W., Stahl, U. and Meyer, V., 2005, New insights into the target site and mode of action of the antifungal protein of Aspergillus giganteus. Res. Microbilol. 156, 47-56, and Theis, T., Wedde, M., Meyer, V. and Stahl, U., 2003, The antifungal protein from Aspergillus giganteus causes membrane permeabilization. Antimicrob. Agents Chemother. 47, 588-593), PAF protein from Penicillium chrysogenum (see Marx, F., Hass, H., Reindl, M., Stoffler, G., Lottspeich, F. and Redl B., 1995, Cloning, structural organization and regulation of expression of the Penicillium chrysogenum paf gene encoding an abundantly secreted protein with antifungal activity. Gene 167, 167-171, and Kaiserer et al., 2003), and Anafp protein from Aspergillus niger (see Lee, G. D., Shin, S. Y., Maeng, C. Y., Jin, Z. Z., Kim, K. L. and Hahm, K S., 1999, Isolation and characterization of a novel antifungal peptide from Aspergillus niger. Biochem. Biophys. Res. Commun. 263, 646-651). The aforementioned antifungal proteins are all secretary proteins, and they can inhibit the growth of a wide range of fungi, but do not influence bacteria and yeasts. These antifungal proteins have similar molecular characteristics, but there are only 42% sequence similarities between the amino acid sequences of the antifungal proteins PAF and AFP (see Kaiserer et al., 2003). These fungus-derived antifungal proteins mainly inhibit fungi of genus Aspergillus and Fusarium spp. (see Theis et al., 2003, and Kaiserer et al., 2003). PAF protein can further inhibit human and animal fungal pathoghes such as Abaidia spp., Mortierella spp., Rhizomucor spp. and Rhizopus spp. These proteins are useful not only as biologically controlling agents for plant fungal pathogen, but also as potential human and animal antifungal drugs (see GalgÓczy, L., Papp. T., Letter, É. Marx, F., Pócsi, I. And Vágvölgyi, C., 2005, Sensitivity of different Zygomycetes to the Penicillium chrysogenum antifungal protein (PAF). J. Basic microbial. 45, 136-141). In addition, it has been reported that the resistance of rice to the rice blast pathogen Magnaporthe grisea can be enhanced by transfecting the cDNA of AFP protein from Aspergillus giganteus into the rice, and therefore the AFP protein can be used in the prevention of rice blast (see Coca, M., Bortolotti, C., Rufat, M., Penas, G., Eritja, R., Tharreau, D., del Pozo A, M., Messeguer, J. and San Segundo, B., 2004, Transgenic rice plants expressing the antifungal AFP protein from Aspergillus giganteus show enhanced resistance to the rice blast fungus Magnaporthe grisea. Plant Mol. Biol. 54, 245-259, and Moreno, A. B., Martinez Del Pozo, A. and San Segundo B. 2006, Biotechnologically relevant enzymes and proteins: Antifungal mechanism of the Aspergillus giganteus AFP against the rice blast fungus Magnaporthe grisea. Appl. Microbiol. Biotechnol. 72(5):883-895).
Paecilomyces variotii and P. lilacinus are the most ubiquitous species of the genus Paecilomyces and also the most frequently involved in human infections. Endophthalmitis and endocarditis are two of the most common infections produced by P.s variotii and P. lilacinus respectively, and have a very bad prognosis. The failure rate of the standard treatment for the infections is about 40%. The future approaches for the treatments will be using combination therapy or developing new classes of antifungal agents (see Ortoneda, M., Capilla, J., Pastor, F. J., Pujol, I., Yustes, C., Serena, C. and Guarro, J. (2004) In vitro interaction of approved and novel drugs against Paecilomyces spp. Antimicrob. Agents Chemother. 48, 2727-2729). Helminthosporium panici is the pathogen of plant ring spot disease. It is an important topic to use biological molecular technique to effectively prevent the fungal infection and decrease the losses caused by the fungal diseases to human health, economical crops and animal husbandry.
Monascus species are important traditional fungi for fermentation in eastern Asia, and are used in the manufacture of fermented products such as alcoholics, fermented red rice (anka), soybean curd (sufu), soybean sauce, and the like. In addition, Monascus species can produce various metabolites and enzymes, such as monacolin K (see Endo, A., Hasumi, K. and Negishi, S. (1985) Monacolins J and L, new inhibitors of cholesterol biosynthesis produced by Monascus rubber. J. Antibiot. (Tokyo) 38(3):420-2), citrinin (see Hajjaj, H., klaebe, A., Goma, G., Blanc, P. J., Barbier, E. and Francois, J. (2000) Medium-chain fatty acids affect citrinin production in the filamentous fungus Monascus rubber, Appl. Environ. Microbiol. 66(3):1120-5), GABA (see Su, Y. C., Wang, J. J., Lin, T. T. and Pan, T. M. (2003) Production of the secondary metabolites gamma-aminobutyric acid and monacolin K by Monascus. J. Ind. Microbiol. Biotechnol. 30(1):41-6), red and yellow pigments (see Carels, M. and Shepherd, D. (1977) The effect of different nitrogen sources on pigment production and sporulation of Monascus species in submerged, shaken culture. Can. J. Microbiol. 23(10): 1360-72, and Tseng, Y. Y., Chen, M. t. and Lin, C. F. (2000) Growth, pigment production and protease activity of Monascus purpureus as affected by salt, sodium nitrite, polyphosphate and various sugars. J. Appl. Microbiol. 88(1):31-7), and protease (see Tsai, M. S., Hseu, T. H. and Shen, Y. S. (1978) Purification and characterization of an acid protease from Monascus kaoliang. Int. J. Protein Res. 12, 293-302), and thus have high potential in the drug developments and the application of industrial enzymes. Among the applications, citrinin is known to have the activity to inhibit the growth of bacteria. However, there is no literature publication regarding the activity of Monascus species to inhibit the growth of fungi. Our earlier whole genome sequencing and decoding project of Monascus mined a possible antifungal protein gene, and therefore it is suggest that Monascus species may possess antifungal activities.